In vitro adventitious shoot regeneration of native spearmint using internodal explants

An in vitro shoot regeneration procedure was developed for native spearmint (Mentha spicata L.) using internodal explants. Shoot regeneration from internodes was evaluated on Murashige and Skoog (MS) media supplemented with individual cytokinins thidiazuron (TDZ), benzylaminopurine (BA), kinetin (KT), or zeatin (ZT) or various pair wise combinations of these. The highest regeneration was achieved by the second internode on a medium containing MS basal salts, B5 vitamins, 10% coconut water, 1.0 mg.L-1 TDZ, 2.5 m.L-1 ZT, and solidified with 0.2% phytagel. Unlike previous protocols this medium does not need sub culturing and produces elongated shoots in 4 weeks, rather than 6 weeks. Maximum number of shoots (36 per explant after 4 weeks) was observed when internodes from 2-week-old stock plants were used as explant source. The shoots were removed and roots were initiated on medium containing MS basal salts, 0.4 mg.L-1 thiamine-HCL, 100 mg.L-1 myo-inositol, 7.5 g.L-1 agar and 0.01 mg.L-1 proportional to-napthaleneacetic acid (NAA) and then plants were transferred to the greenhouse 2 weeks after root initiation, where 100% of the plantlets developed into healthy plants.